Western blot abreview for Anti-Rad50 antibody [13B3/2C6]

Good
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (lung and foreskin fibroblast cells)
Specification
lung and foreskin fibroblast cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Other product details

Dilution
1/1000
Incubation time
2 hour(s) and 0 minute(s)

Secondary antibody

Name
Non-Abcam antibody was used: Goat Anti-Mouse HRP Conjugated
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase

Detection

Detection method
ECL+
Exposure
30 second(s)
Bands
Specific: 150 kDa
Positive control
whole cell samples
Negative control
(none)

Additional data

Additional Notes
7.5% western blot gels were loaded with 70ug of both IMR-90 and HCA2 whole cell extracts. The samples had been prepared in Laemmlie buffer and a protease inhibitor cocktail. The antibody was then used in a 1/1000 dilution with a monoclonal goat anti-mouse secondary antibody in a 1/5000 dilution. The antibody worked very well for the HCA2 cells. With a 2 hour incubation in the primary antibody, the resulting picture was a nice strong band at 150 kDa (30 sec exposure). The band is a bit smeared underneath, but this does not get in the way of observing the band. There were not any non-specific bands. The band that appeared in the IMR-90 cells was much weaker, and may be impoved by loading more protein on the gel. Overall I would use this antibody again to study Rad50.

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Submitted Mar 02 2006

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